• Title/Summary/Keyword: co-cultures

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Optimal Conditions for Chitinase Production by Serratia marcescens

  • Cha, Jin-Myeong;Cheong, Kyung-Hoon;Cha, Wol-Suk;Choi, Du-Bok;Roh, Sung-Hee;Kim, Sun-Il
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.4
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    • pp.297-302
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    • 2004
  • A chitinase-producing bacterium was isolated from seashore mud around Beobseongpo in Chunmam province through the use of a selective enrichment culture. The best chitinase producing strain was isolated and identified as Serratia marcescens KY from its characteristics. For effective production of chitinase, optimum pH, temperature, and agitation speed were investigated in flask cultures. The optimum pH using Serratia marcescens KY was between pH 6 and 7 and the chitinase produced was 37.9 unit/mL. On the other hand, the optimal pH of the Serratia marcescens ATCC 27117 was 7.5, and the produced amount of chitinase was 35.2 unit/mL. The optimal temperature for chitinase production for Serratia marcescens KY and Serratia marcescens ATCC 27117 was $30^{\circ}$. The cell growth pattern at different temperature was almost identical to the chitinase production. To investigate the optimal shaking speed under optimal culture, speeds were varied in the range of 0∼300 rpm. The maximum production of chitinase was carried at 200 rpm although the cell growth was the highest at 150 rpm. It indicates that oxygen adjustment is required for the maximum chitinase production. Using optimal conditions, batch cultures for comparing Serratia marcescens KY and Serratia marcescens ATCC 27117 were carried out in a 5 L fermentor. The oxygen consumption was increased with the increase of culture. Especially, at 120 h of culture Serratia marcescens KY and Serratia marcescens ATCC 27117 produced 38.3 unit/mL, and 33.5 unit/mL, respectively.

Characterization and Anti-Helicobactor pylori Activity of Xanthium strumarium L. Extract on Lactic acid Fermentation (도꼬마리 추출물의 유산발효 특성 및 Helicobactor pylori에 대한 항균활성)

  • Kang, Dong-Hee;Kim, Hyun-Soo
    • KSBB Journal
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    • v.25 no.3
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    • pp.244-250
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    • 2010
  • This study characterized the anti-Helicobactor pylori activity of Xanthium strumarium L. extract obtained by lactic acid fermentation. The growth of the Lactobacillus strains was typically robust upon lactic acid production in monocultures containing Xanthium strumarium L. extract. Lactic acid fermentation in mixed cultures of Lactobacillus brevis KCTC 3498 and Lactobacillus casei KCTC 3109 produced higher of anti- H. pylori activity than monocultures. Concerning antioxidant activity of fermented extracts, total polyphenol contents were the highest in co-cultures of Lactobacillus brevis KCTC 3498 and Lactobacillus helveticus KCTC 3545. Electron donating ability using diphenyl picryl hydrazyl (DPPH) showed 70% scavenging in Lactobacillus casei KCTC 3109.

Effective Application of Multiplex RT-PCR for Characterization of Human Embryonic Stem Cells/ Induced Pluripotent Stem Cells (다중 역전사 중합효소 연쇄 반응(Multiplex RT-PCR)을 이용한 인간배아 줄기세포 및 유도만능 줄기세포의 효과적인 분화 양상 조사)

  • Kim, Jung-Mo;Cho, Youn-Jeong;Son, On-Ju;Hong, Ki-Sung;Chung, Hyung-Min
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.1-8
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    • 2011
  • Techniques to evaluate gene expression profiling, such as sufficiently sensitive cDNA microarrays or real-time quantitative PCR, are efficient methods for monitoring human pluripotent stem cell (hESC/iPSC) cultures. However, most of these high-throughput tests have a limited use due to high cost, extended turn-around time, and the involvement of highly specialized technical expertise. Hence, there is an urgency of rapid, cost-effective, robust, yet sensitive method development for routine screening of hESCs/hiPSCs. A critical requirement in hESC/hiPSC cultures is to maintain a uniform undifferentiated state and to determine their differentiation capacity by showing the expression of gene markers representing all three germ layers, including ectoderm, mesoderm, and endoderm. To quantify the modulation of gene expression in hESCs/hiPSC during their propagation, expansion, and differentiation via embryoid body (EB) formation, we developed a simple, rapid, inexpensive, and definitive multimarker, semiquantitative multiplex RT-PCR platform technology. Among the 9 gene primers tested, 5 were pluripotent markers comprising set 1, and 3 lineage-specific markers were combined as set 2, respectively. We found that these 2 sets were not only effective in determining the relative differentiation in hESCs/hiPSCs, but were easily reproducible. In this study, we used the hES/hiPS cell lines to standardize the technique. This multiplex RT-PCR assay is flexible and, by selecting appropriate reporter genes, can be designed for characterization of different hESC/hiPSC lines during routine maintenance and directed differentiation.

Bead-to-Bead Cell Transfer by Induction of Detachment of Anchorage Dependent HeLa Cells Grown on Macroporous Microcarriers (부착성 HeLa 세포의 탈리 유도에 의한 다공성 미립담체의 담체간 전이 배양)

  • 이두훈;박정극
    • KSBB Journal
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    • v.13 no.1
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    • pp.83-89
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    • 1998
  • Using a cellulose macroporous microcarrier, HeLa cells were cultivated in 100mL spinner flask(Bellco Co., USA) and confluent cell laden microcarriers were subcultured by bead-to-bead cell transfer method. In macroporous mcirocarrier-HeLa system viable suspended cells played an important role in bead-to-bead cell transfer and that could be increased by use of RPMI-1640, a calcium-ion-reduced-media and high speed agitation. Successful bead-to-bead cell transfers were performed continuously three time in spinner flask. We applied this technique to produce recombinant Vaccinia virus which express $\beta$-galactosidase. Recombinant protein yield of bead-to-bead transferred culture was comparable to conventional microcarrier cultures that were inoculated by cells detached from T-flask. Although trypsinization is a useful method for subculturing microcarriers in some cases, that process adds quality control problem and handling steps to large scale cell production. There fore, bead-to-bead cell transfer technique offers another convenient and efficient scale-up method for continuous microcarrier cultures.

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A Study on the Spatial Transformation through the Process of Spatial Changes of Dining-Kitchen for Japanese Houses in Korea (부엌의 변화과정을 통해 본 일식주택 공간적 변화에 관한 연구)

  • Lee, So-Jin;Kim, Young-Bum;Park, Yong-Hwan
    • Journal of the Korean housing association
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    • v.19 no.3
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    • pp.83-94
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    • 2008
  • When Korea opened its ports and underwent Japanese colonization, many Japanese style houses were built in Korea. Following Korea's independence from Japan, Koreans began to reside in these houses. The objective of this study is to examine the current state of Japanese style residence areas and Japanese style houses in Korea, and to determine the change in the characteristics of dining kitchens that have taken place since Koreans have lived in them. In the process, while assimilation occurred, there was also a conflict between the residential lifestyles of the two cultures, developing into a state where two housing cultures co-existed. The dining kitchens showed the most sensitive adjustments to social changes, facilitating a number of important changes in the process of modernizing houses. In this regard, the intention is to determine how the dining kitchens responded to other areas within the house as they were being transformed. Research for this study is based on previous studies that were carried out in 1991 on Japanese style houses, in order to clearly define the process of change chronologically rather than from a single examination. In consequently, From the process of changes where from a conventional kitchen to DK anger, 1) The public space - wooden floor, living room, etc - had been formed in house spaces. 2) In the lifestyle, privacy secured. It was separated each functional spaces that greeting space for guests and family's space in the lifestyle. 3) The cause of variation could be summarized that differences of living style, a change of life and fuel.

Enhancement of centelloside production from cultured plants of Centella asiatica by combination of thidiazuron and methyl jasmonate

  • Yoo, Nam-Hee;Kim, Ok-Tae;Kim, Jung-Bong;Kim, Sun-Hee;Kim, Young-Chang;Bang, Kyong-Hwan;Hyun, Dong-Yun;Cha, Seon-Woo;Kim, Min-Young;Hwang, Baik
    • Plant Biotechnology Reports
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    • v.5 no.3
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    • pp.283-287
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    • 2011
  • In order to produce centellosides from whole plant cultures of Centella asiatica (L.) Urban, we evaluated the synergistic effects of thidiazuron (TDZ) and methyl jasmonate (MJ) on whole plant growth and centelloside production. After 4 weeks of treatment with 0.025 mg/L of TDZ coupled with 0.1 mM MJ, the production of made-cassoside and asiaticoside from whole plant cultures was estimated to be 2.40- and 2.44-fold, respectively, above that of MJ elicitation alone. When whole plants were treated with a growth regulator and an elicitor, the growth of whole plants, as compared to the controls, did not differ. Additionally, total phytosyterol content in the leaves of whole plants co-treated with MJ and TDZ was 1.08-fold greater than those of MJ alone. These results demonstrate that combined treatments not only stimulate the accumulation of centellosides in the leaves but also inhibit the reduction of phytosterol levels caused by MJ elicitation.

Absorbance Spectrum for Mesodinium rubrum MR-MAL01, a marine photosynthetic ciliate, fed on Photo-adapted Cryptophyte (광적응된 은편모조류를 섭식한 해양 광합성 섬모류 Mesodinium rubrum MR-MAL01의 흡광스펙트럼)

  • 김형섭;명금옥;조수근;이원호
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.8 no.1
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    • pp.29-34
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    • 2003
  • Recent reports on the phagotrophic feeding of M. rubrum are based on cultivation experiments with novel isolates of this ciliate species from Gomso Bay, Korea. Photo-adapted cryptophyte(CR-MAL01) cultures at high light of 100 $\mu$mol photons m$^{-2}$ s$^{-1}$ (HL) and low light of 10 $\mu$mol photons m$^{-2}$ s$^{-1}$ (LL) were fed to M. rubrum (MR-MAL01) cultures under HL and LL conditions, respectively. Absorbance spectrum by LL M. rubrum showed the same peak at wavelengths around 542nm as that by LL cryptophyte prey, which was not showed in HL M. rubrum. This result supports the implication that light utilization and absorption pattern of M. rubrum population must depend on the status of photo-adaptation of the co-existing population of prey cryptophyte. Consequences of the present research results were discussed in relation to the function of the prey cryptophyte and phagotrophic M. rubrum in marine microbial ecosystem.

The Influences of Extracts from Ceratium spp, on the Growth of Harmful Microalgae (유해성 적조생물에 대한 Ceratium 추출물 영향 평가)

  • Cho, Eun-Seob
    • Journal of Life Science
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    • v.14 no.4
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    • pp.589-592
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    • 2004
  • The growth response of the fish-killing dinoflagellate Cochlodinium polykrikoides was studied in cultures, using the treatment of Ceratium extracts by a methanol, a water-soluble, and a cell-free medium. The cell-free medium had the most increasing on the growth of C. polykrikoides cultures, enriched with $\geq$ 25% Ceratium, whereas the methanol and water-souble fractions did not affect the growth of C. polykrikoides exposed to even higher concentration. In particular, the cell-free medium also increased the growth of Gyrodinium impudicum and Chaetoceros sp., similar species to C. polykrikoides. In contrast to C. polykrikoides, G. impudicum and Chaetoceros sp., the growth of Alexandrium tamarense was inhibited significantly, and there was no great effect on the growth of Prorocentrum minimum. These results imply that Ceratium extracts may play an important role in the stimulatory effect of C. polykrikoides, and they have to affect the interaction between C. polykrikoides and Ceratium when co-existing.

A newly developed consensus polymerase chain reaction to detect Mycoplasma species using 16S ribosomal RNA gene

  • Hong, Sunhwa;Park, Sang-Ho;Chung, Yung-Ho;Kim, Okjin
    • Korean Journal of Veterinary Service
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    • v.35 no.4
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    • pp.289-294
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    • 2012
  • Mycoplasmas are highly fastidious bacteria, difficult to culture and slow growing. Infections with Mycoplasma species can cause a variety of problems in living organisms and in vitro cell cultures. In this study, we investigated the usefulness of a genus-specific consensus PCR analysis method to detect Mycoplasma species. The developed consensus primer pairs MycoF and MycoR were designed specifically to amplify the 16S ribosomal RNA gene (rRNA) of Mycoplasma species by the optimized PCR system. The developed consensus PCR system effectively amplified 215 bp of Mycoplasma genus-specific region of 16S rRNA. In conclusion, we recommend this consensus PCR for monitoring Mycoplasma species in animals, human and cell culture system.

The Effect of Light on the Production of Reserpine in Cultured Rauwolfia serpentina Cells

  • Yamamoto, Osamu
    • Natural Product Sciences
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    • v.2 no.2
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    • pp.90-95
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    • 1996
  • When reserpine-producing cell strains of Rauwolfia serpentina were transferred from the dark to the light irradiation, the production of reserpine was extremely enhanced whereas the cell growth was suppressed. In an incubation period of 20 days, the most effective culture condition for reserpine production was the combination of 8 days of dark culture and following 12 days of light culture. The time courses of both cell growth and reserpine production were measured in vitro in order to clarify the effect of wave length range of light on the biosynthesis of reserpine. Although the growth of cultured cells which had been incubated under continuous red, yellow, and green lights, respectively, was similar to that of the cultured cells subcultured in the dark. The cells cultured under red light irradiation produced less reserpine than dark-grown cultures. Both blue and near-ultraviolet light inhibited the growth of cultured cells. The production of reserpine was strikingly enhanced by blue light, but was strongly inhibited by near-ultraviolet light.

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